Publications

In this study, we compared multiple quantitative ultrasound metrics for the purpose of differentiating muscle in 20 healthy, 10 dystrophic and 10 obese mice. High-frequency ultrasound scans were acquired on dystrophic (D2-mdx), obese (db/db) and control mouse hindlimbs. A total of 248 image features were extracted from each scan, using brightness-mode statistics, Canny edge detection metrics, Haralick features, envelope statistics and radiofrequency statistics. Naïve Bayes and other classifiers were trained on single and pairs of features. The a parameter from the Homodyned K distribution at 40 MHz achieved the best univariate classification (accuracy = 85.3%). Maximum classification accuracy of 97.7% was achieved using a logistic regression classifier on the feature pair of a2 (K distribution) at 30 MHz and brightness-mode variance at 40MHz. Dystrophic and obese mice have muscle with distinct acoustic properties and can be classified to a high level of accuracy using a combination of multiple features.

Fibrodysplasia ossificans progressiva (FOP) is an ultra-rare disorder involving skeletal dysplasia and heterotopic ossification (HO) of muscle and connective tissue. We aimed to define a novel biomarker in FOP that enables reliable assessment of musculoskeletal tissue integrity. Considering logistical difficulties that FOP patients often face, our goal was to identify an at-home biomarker technique. Electrical impedance myography (EIM) is a non-invasive, portable method that can inform on muscle health. 15 FOP patients (age 10-52) and 13 healthy controls were assessed. Using EIM, multiple muscle groups were characterized per participant in a 45-min period. The Cumulative Analogue Joint Involvement Scale (CAJIS) was implemented to determine mobility burden severity. We additionally evaluated physical activity levels via a Patient-Reported Outcomes Measurement Information System (PROMIS)-based questionnaire. Relative to controls, FOP patients demonstrated significantly lower regional and whole-body phase values at 50 kHz and 100 kHz, indicating more diseased muscle tissue. Lower whole-body phase and reactance values, and higher resistance values, were associated with greater FOP burden (CAJIS score range: 4-30) and lower physical activity levels at 50 kHz and 100 kHz. This study points to the potential utility of EIM as a clinical biomarker tool capable of characterizing muscle integrity in FOP.

To aid in the identification of new treatments for amyotrophic lateral sclerosis (ALS), convenient biomarkers are needed to effectively and uniformly measure drug efficacy. To this end, we assessed the effects of the nutraceutical resveratrol (RSV) on disease onset and overall survival in SOD1-G93A (ALS) mice and compared several standard biomarkers including body mass, motor score (MS), paw grip endurance (PGE), and compound motor action potential (CMAP) amplitude, with the technique of electrical impedance myography (EIM) to follow disease progression. Eighteen ALS mice (nine females, nine males) received RSV in the chow (dose: 120 mg/kg/day) starting at 8 weeks of age; 19 ALS mice (nine females, 10 males) received normal chow; 10 wild type (WT) littermates (five females, five males) fed standard chow served as controls. Biomarker assessments were performed weekly beginning at 8 weeks. No differences in either disease onset or overall survival were found between RSV-treated and untreated ALS mice of either sex; moreover, all biomarkers failed to identify any beneficial effect of RSV when administered at this dose. Therefore, for the comparative evaluation of the ability of the various biomarkers to detect the earliest symptoms of disease, data from all animals (i.e., RSV-treated and untreated ALS mice of both sexes) were combined. Of the biomarkers tested, EIM impedance values, i.e., surface EIM longitudinal phase at 50 kHz (LP 50 kHz), and CMAP amplitude showed the earliest significant changes from baseline. LP 50 kHz values showed a rate of decline equivalent to that of CMAP amplitude and correlated with both PGE and CMAP amplitude [Spearman rho = 0.806 (p = 0.004) and 0.627 (p = 0.044), respectively]. Consistent with previous work, these findings indicate that surface EIM can serve as an effective non-invasive biomarker for preclinical drug testing in rodent models of ALS.

Convenient tools to assess canine skeletal muscle health would be useful for a variety of applications, including standard veterinary assessments of dog fitness, as well as studies of muscle deterioration due to age or disease. One technology that can be applied conveniently to awake dogs with minimal restraint is electrical impedance myography (EIM). In EIM, a weak electrical current is applied via surface electrodes to a muscle of interest and consequent impedance characteristics of the muscle are obtained, providing insight into muscle condition and composition. In this study, we assessed a total of 73 dogs (42 males and 31 females), of varied neutering status and breed, ages 0.6 to 13.5 years. We identified age-dependent reference values for the 100 kHz phase value in three pelvic limb muscles, caudal sartorius, cranial tibial, and gastrocnemius. While phase values were generally higher in males than females, the difference did not reach significance. In general, values declined on average with age at about 0.5 degrees/year, but with the decline being most substantial in the oldest dogs. Limited reproducibility assessment of the technique suggested good repeatability with variation in values between measurements being under 5%. These results show that EIM has the potential for the assessment of canine muscle health and may find value in aging muscle research.

OBJECTIVE: The Cole model is a widely used fractional circuit model in electrical bioimpedance applications for evaluating the content and status of biological tissues and fluids. Existing methods for estimating the Cole impedance parameters are often based on multi-frequency data obtained from stepped-sine measurements fitted using a complex non-linear least square (CNLS) algorithm. Newly emerged numerical methods from the magnitude of electrical bio-impedance data-only do not need CNLS fitting, but they still require multi-frequency stepped-sine data. This study proposes a novel approach to estimating the Cole impedance parameters that combines a numerical and time-domain fitting method based on a single-frequency DC-biased sinusoidal current excitation.

APPROACH: First, the transient and steady-state voltage response along with the current excitation are acquired in electrical bio-impedance measurement. From the sampled data, a numerical method is applied to provide the initial estimation of the Cole impedance parameters, which are then used in a time-domain iterative fitting algorithm.

RESULTS: The accuracy of the algorithm proposed is tested with noisy electrical bio-impedance simulations. The maximum relative error of the estimated Cole impedance parameters is 1% considering 2% (34 dB) additive Gaussian noise. Experimental measurements performed on a 2R-1C circuit and some fruit samples show a mean difference less than 1% and 5% respectively compared to the Cole impedance parameters estimated from a commercial electrical bio-impedance analyzer performing stepped-sine measurements and CNLS fitting.

SIGNIFICANCE: This is the first method that allows estimating the Cole impedance parameters from single-frequency electrical bio-impedance data. The approach presented could find broad use in many applications, including single-frequency body impedance analysis.

INTRODUCTION: Surface electrical impedance myography (sEIM) has the potential for providing information on muscle composition and structure noninvasively. We sought to evaluate its use to predict myofiber size and connective tissue deposition in the D2-mdx model of Duchenne muscular dystrophy (DMD).

METHODS: We applied a prediction algorithm, the least absolute shrinkage and selection operator, to select specific EIM measurements obtained with surface and ex vivo EIM data from D2-mdx and wild-type (WT) mice (analyzed together or separately). We assessed myofiber cross-sectional area histologically and hydroxyproline (HP), a surrogate measure for connective tissue content, biochemically.

RESULTS: Using WT and D2-mdx impedance values together in the algorithm, sEIM gave average root-mean-square errors (RMSEs) of 26.6% for CSA and 45.8% for HP, which translate into mean errors of ±363 μm2 for a mean CSA of 1365 μm2 and of ±1.44 μg HP/mg muscle for a mean HP content of 3.15 μg HP/mg muscle. Stronger predictions were obtained by analyzing sEIM data from D2-mdx animals alone (RMSEs of 15.3% for CSA and 34.1% for HP content). Predictions made using ex vivo EIM data from D2-mdx animals alone were nearly equivalent to those obtained with sEIM data (RMSE of 16.59% for CSA), and slightly more accurate for HP (RMSE of 26.7%).

DISCUSSION: Surface EIM combined with a predictive algorithm can provide estimates of muscle pathology comparable to values obtained using ex vivo EIM, and can be used as a surrogate measure of disease severity and progression and response to therapy.

Pathological age-related loss of skeletal muscle strength and mass contribute to impaired physical function in older adults. Factors that promote the development of these conditions remain incompletely understood, impeding development of effective and specific diagnostic and therapeutic approaches. Inconclusive evidence across species suggests disruption of action potential signal transmission at the neuromuscular junction (NMJ), the crucial connection between the nervous and muscular systems, as a possible contributor to age-related muscle dysfunction. Here we investigated age-related loss of NMJ function using clinically relevant, electrophysiological measures (single-fiber electromyography (SFEMG) and repetitive nerve stimulation (RNS)) in aged (26 months) versus young (6 months) F344 rats. Measures of muscle function (e.g., grip strength, peak plantarflexion contractility torque) and mass were assessed for correlations with physiological measures (e.g., indices of NMJ transmission). Other outcomes also included plantarflexion muscle contractility tetanic torque fade during 1-s trains of stimulation as well as gastrocnemius motor unit size and number. Profiling NMJ function in aged rats identified significant declines in NMJ transmission stability and reliability. Further, NMJ deficits were tightly correlated with hindlimb grip strength, gastrocnemius muscle weight, loss of peak contractility torque, degree of tetanic fade, and motor unit loss. Thus, these findings provide direct evidence for NMJ dysfunction as a potential mechanism of age-related muscle dysfunction pathogenesis and severity. These findings also suggest that NMJ transmission modulation may serve as a target for therapeutic development for age-related loss of physical function.

Skeletal muscle function deficits associated with advancing age are due to several physiological and morphological changes including loss of muscle size and quality (conceptualized as a reduction in the intrinsic force-generating capacity of a muscle when adjusted for muscle size). Several factors can contribute to loss of muscle quality, including denervation, excitation-contraction uncoupling, increased fibrosis, and myosteatosis (excessive levels of inter- and intramuscular adipose tissue and intramyocellular lipids). These factors also adversely affect metabolic function. There is a major unmet need for tools to rapidly and easily assess muscle mass and quality in clinical settings with minimal patient and provider burden. Herein, we discuss the potential for electrical impedance myography (EIM) as a tool to evaluate muscle mass and quality in older adults. EIM applies weak, non-detectible (e.g., 400 μA), mutifrequency (e.g., 1 kHz-1 MHz) electrical currents to a muscle (or muscle group) through two excitation electrodes, and resulting voltages are measured via two sense electrodes. Measurements are fast ( 5 s/muscle), simple to perform, and unaffected by factors such as hydration that may affect other simple measures of muscle status. After nearly 2 decades of study, EIM has been shown to reflect muscle health status, including the presence of atrophy, fibrosis, and fatty infiltration, in a variety of conditions (e.g., developmental growth and maturation, conditioning/deconditioning, and obesity) and neuromuscular diseases states [e.g., amyotrophic lateral sclerosis (ALS) and muscular dystrophies]. In this article, we describe prior work and current evidence of EIM's potential utility as a measure of muscle health in aging and geriatric medicine.